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目的建立快速分子指纹图谱分析法,应用于都柏林念珠菌和白念珠菌分子指纹图谱差异性分析。方法以野生株噬菌体M13微卫星特异核心序列为单一引物,分别进行都柏林念珠菌和白念珠菌基因组DNA的聚合酶链反应(PCR)扩增,并将产生的分子指纹图谱分别应用琼脂糖凝胶电泳和微流DNA芯片分析。结果17株都柏林念珠菌和白念珠菌基因组DNA的扩增产物经琼脂糖凝胶电泳分析,见都柏林念珠菌和白念珠菌各有5条固定阳性条带出现,两者之间阳性条带大小差异有统计学意义。微流DNA芯片显示都柏林念珠菌固定出现阳性条带大小依次为960、1177、1297、1495、1797bp;白念珠菌固定出现阳性条带大小依次为653、1323、1531、2021、2875bp。结论都柏林念珠菌和白念珠菌分子指纹图谱存在显著差别,可应用于两者的快速鉴别,尤其是微流DNA芯片法简便、快速、重复性好,更适合于大规模分子流行病学监测。
Objective To establish a rapid molecular fingerprinting method for the differential analysis of molecular fingerprinting of Candida albicans and Candida albicans in Dublin. Methods The M13 microsatellite-specific core sequence of phage M13 was used as a single primer to amplify the genomic DNA of Candida albicans and Candida albicans, respectively. The molecular fingerprints of the strains were amplified by agarose gel Electrophoresis and microfluidic DNA chip analysis. Results The amplification products of 17 strains of Candida albicans and Candida albicans genomic DNA were analyzed by agarose gel electrophoresis. There were 5 fixed positive bands in Candida albicans and Candida albicans respectively. The positive bands between them The difference was statistically significant. Microfluidic DNA chips showed positive bands of Candida albicans fixation appeared in order of 960,1177,1297,1495,1797bp; Candida albicans positive bands appeared in the order of 653,1323,1531,2021,2875bp. Conclusion There are significant differences between fingerprints of Candida albicans and Candida albicans, which can be used for the rapid identification of the two. Especially the microfluidic DNA chip method is simple, rapid and reproducible, and is more suitable for large-scale molecular epidemiological surveillance.